title: High frequency of de novo mutations in ankyrin gene (ANK1) in children with hereditary spherocytosis.
abstract: OBJECTIVE
  To evaluate the frequency of de novo monoallelic expression of the ANK1 gene in hereditary spherocytosis individuals appearing as recessive.
  
  
  METHODS
  We studied 40 unrelated children with spherocytosis and their normal parents. The genomic distribution of the ankyrin (AC)n dinucleotide repeats was evaluated in the patients showing combined ankyrin and spectrin deficiency. To search for the absence of mRNA derived from one of the two ANK1 genes, cDNA from the heterozygous patients was amplified using polymerase chain reaction. This was analyzed for the (AC)n dinucleotide repeats.
  
  
  RESULTS
  Thirty-three hereditary spherocytosis subjects had variable degrees of combined ankyrin and spectrin reduction; 19 were found to be heterozygous for the AC repeat lengths and were further studied. In 12, we found a cDNA polymerase chain reaction product from one ankyrin gene alone. These findings strongly suggested the nonexpression of one of the two ANK1 genes because of the de novo mutational events.
  
  
  CONCLUSIONS
  The de novo loss of an ankyrin allele expression is a frequent cause of hereditary spherocytosis in children with normal parents. Therefore the category of genuinely recessive hereditary spherocytosis cases is further reduced compared with spherocytosis cases because of de novo mutations. The determination of the (AC)n microsatellite polymorphisms appears as a helpful and reliable tool for the discrimination between these two categories.
authors.affiliation: Department of Pediatrics, Second University of Napoli, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Miraglia del Giudice
authors.forename: M
authors.initials: M
authors.lastname: Francese
authors.forename: B
authors.initials: B
authors.lastname: Nobili
authors.forename: L
authors.initials: L
authors.lastname: Morlé
authors.forename: S
authors.initials: S
authors.lastname: Cutillo
authors.forename: J
authors.initials: J
authors.lastname: Delaunay
authors.forename: S
authors.initials: S
authors.lastname: Perrotta
mesh_terms: D000293:Adolescent; D017487:Ankyrins / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D018900:Dinucleotide Repeats; D005260:Female; D005808:Genes, Recessive; D006801:Humans; D007223:Infant; D008297:Male; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D013103:Spherocytosis, Hereditary / Q000235:genetics*
chemical_list: C110202:ANK1 protein, human; D017487:Ankyrins; D012333:RNA, Messenger